| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Papers In Press, published online ahead of print April 1, 2002
Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, CT 06520-8026
Corresponding Author: catherine.berlot{at}yale.edu
To investigate the subcellular organization of receptor-G protein signaling pathways, a robust dominant negative
J. Biol. Chem, 10.1074/jbc.M201330200
Submitted on February 8, 2002
Revised on March 27, 2002
Accepted on April 1, 2002
A highly effective dominant negative
s construct containing mutations that affect distinct functions inhibits multiple Gs-coupled receptor signaling pathways
s mutant containing substitutions that alter distinct functions was produced and tested for its effects on Gs-coupled receptor activity in HEK-293 cells. Mutations in the 3
5 loop region, which increase receptor affinity, decrease receptor-mediated activation, and impair activation of adenylyl cyclase, were combined with G226A, which increases affinity for 
, and A366S, which decreases affinity for GDP. This triple s mutant can inhibit signaling to Gs from the luteinizing hormone receptor by 97% and from the calcitonin receptor by 100%. In addition, this s mutant blocks all signaling from the calcitonin receptor to Gq. These results lead to two conclusions about receptor-G protein signaling. First, individual receptors have access to multiple types of G proteins in HEK-293 cell membranes. Second, different G protein subunits can compete with each other for binding to the same receptor. This dominant negative s construct will be useful for determining interrelationships among distinct receptor-G protein interactions in a wide variety of cells and tissues.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  A. R. Zurita and L. Birnbaumer The same mutation in Gs{alpha} and transducin {alpha} reveals behavioral differences between these highly homologous G protein {alpha}-subunits PNAS, February 19, 2008; 105(7): 2363 - 2368. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Dean, J.-P. Vilardaga, J. T. Potts Jr., and T. J. Gardella Altered Selectivity of Parathyroid Hormone (PTH) and PTH-Related Protein (PTHrP) for Distinct Conformations of the PTH/PTHrP Receptor Mol. Endocrinol., January 1, 2008; 22(1): 156 - 166. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Nakamura, B. Han, T. Nishishita, Y. Bai, and K. Kakudo Calcitonin targets extracellular signal-regulated kinase signaling pathway in human cancers J. Mol. Endocrinol., December 1, 2007; 39(6): 375 - 384. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. G. Chepurny and G. G. Holz A Novel Cyclic Adenosine Monophosphate Responsive Luciferase Reporter Incorporating a Nonpalindromic Cyclic Adenosine Monophosphate Response Element Provides Optimal Performance for Use in G Protein Coupled Receptor Drug Discovery Efforts J Biomol Screen, August 1, 2007; 12(5): 740 - 746. [Abstract] [PDF]
|
|
|
|
 |
|
 S. Tanaka, K. Ishii, K. Kasai, S. O. Yoon, and Y. Saeki Neural Expression of G Protein-coupled Receptors GPR3, GPR6, and GPR12 Up-regulates Cyclic AMP Levels and Promotes Neurite Outgrowth J. Biol. Chem., April 6, 2007; 282(14): 10506 - 10515. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Lariviere, G. Garrel, V. Simon, J.-W. Soh, J.-N. Laverriere, R. Counis, and J. Cohen-Tannoudji Gonadotropin-Releasing Hormone Couples to 3',5'-Cyclic Adenosine-5'-Monophosphate Pathway through Novel Protein Kinase C{delta} and -{epsilon} in L{beta}T2 Gonadotrope Cells Endocrinology, March 1, 2007; 148(3): 1099 - 1107. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Dean, A. Khatri, Z. Potetinova, G. E. Willick, and T. J. Gardella Role of Amino Acid Side Chains in Region 17-31 of Parathyroid Hormone (PTH) in Binding to the PTH Receptor J. Biol. Chem., October 27, 2006; 281(43): 32485 - 32495. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Pereira and R. A. Cerione A Switch 3 Point Mutation in the {alpha} Subunit of Transducin Yields a Unique Dominant-negative Inhibitor J. Biol. Chem., October 21, 2005; 280(42): 35696 - 35703. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. A. Vortherms, C. H. Nguyen, C. H. Berlot, and V. J. Watts Using Molecular Tools to Dissect the Role of G{alpha}s in Sensitization of AC1 Mol. Pharmacol., December 1, 2004; 66(6): 1617 - 1624. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. H. Cleator, R. Ravenell, D. T. Kurtz, and J. D. Hildebrandt A Dominant Negative G{alpha}s Mutant That Prevents Thyroid-stimulating Hormone Receptor Activation of cAMP Production and Inositol 1,4,5-Trisphosphate Turnover: COMPETITION BY DIFFERENT G PROTEINS FOR ACTIVATION BY A COMMON RECEPTOR J. Biol. Chem., August 27, 2004; 279(35): 36601 - 36607. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y.-L. Wu, S. B. Hooks, T. K. Harden, and H. G. Dohlman Dominant-negative Inhibition of Pheromone Receptor Signaling by a Single Point Mutation in the G Protein {alpha} Subunit J. Biol. Chem., August 20, 2004; 279(34): 35287 - 35297. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. A. Roy, K. E. Lemberg, and P. Chidiac Recruitment of RGS2 and RGS4 to the Plasma Membrane by G Proteins and Receptors Reflects Functional Interactions Mol. Pharmacol., September 1, 2003; 64(3): 587 - 593. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
